Recombinant human IFN-α6 Mammalian cell-expressed human interferon alpha 6 (alpha K) with HSA Cat. code: rcyc-hifna6 https://www.invivogen.com/human-ifna6 For research use only, not for diagnostic or therapeutic use Version 18I12-NJ PRODUCT INFORMATION BACKGROUND Contents Type I interferons (IFN) include the IFN-α family, IFN-β, IFN-ε, IFN-κ • 1 µg of lyophilized recombinant human IFN-α6 and IFN-ω. IFN-αs are important anti-viral cytokines that also play Note: This product is sterile filtered prior to lyophilization. anti-proliferative and immuno-modulatory functions. The human IFN-α • 1.5 ml endotoxin-free water family comprises 13 genes encoding 12 proteins, IFN-α13 being identical to IFN-α1. All IFN-αs bind to a common heterodimer receptor IFNAR1/ Storage and stability IFNAR2. The ternary complex signals through the Janus kinase (JAK) • Recombinant human IFN-α6 is shipped at room temperature. Upon and signal transducer and activator of transcription (STAT) signaling receipt it should be immediately stored at -20 °C. Lyophilized product is stable for 6 months at -20°C. pathway, inducing the formation of the ISGF3 transcriptional complex • Upon resuspension, prepare aliquots of recombinant human IFN-α6 and (STAT1/STAT2/IRF9). ISGF3 binds to IFN-stimulated response elements 2 store at -80 °C for 4 months. (ISRE) in the promoters of numerous IFN-stimulated genes (ISGs) . Note: Avoid repeated freeze-thaw cycles. Quality control IFN-α IFN-α • Purity: ≥ 95% (SDS-PAGE) Low affinity High affinity • Endotoxin: ≤ 1 EU/µg • The biological activity has been confirmed using HEK-Blue™ IFN-α/β IFNAR1 IFNAR2 cells (see validation data sheet available on our website). TyK2 JAK1 Other factors • The units have been determined for each lot using hIFN-α2 laboratory e.g. STAT2/STAT6, standard calibrated to the WHO International Standard Interferon Alpha Crkl/STAT5 (Human leukocyte-derived) from the National Institute for Biological 1 STAT2 Standards and Control (NIBSC; code 94/784) . 1 2 ISGF3 TAT TAT 1. http://www.nibsc.org/documents/ifu/94-784.pdf. National Institute for Biological complex } S S STAT6 Standards and Control. Potters Bar, Hertfordshire, EN6 3QG. WHO International IRF9 Crkl Laboratory for Biological Standards. UK Official Medicines Control Laboratory. STAT5 Tunable ISGs CHARACTERISTICS Robust ISGs (anǎ-proliferaǎve, ISRE (anǎ-viral) GAS Source: Mammalian; Chinese hamster ovary (CHO) cells ISRE immuno-modulatory) Molecular mass: ≈ 22 kDa (SDS-PAGE) Robust and tunable ISGs induction by IFN-αs. Chromosome: 9 UniProt ID: P05013 Human IFN-α genes have evolved under strong selective pressure, Alternate name: IFN-alpha K suggesting a non-redundant role between IFN-α subtypes3. Although Formulation: Recombinant human IFN-α6 was lyophilized from a most studies have focused on IFN-α2 and IFN-α8, a consensus model for 0.2 µm filtered phosphate buffer solution (pH 7.4) containing 2% human all IFN-αs has emerged depending on the affinity of a particular IFN-α serum albumin (HSA) and 5% saccharose. subtype for IFNAR. Low affinity IFN-α subtypes signal strictly through Solubility: 100 µg/ml in water ISGF3 and induce robust ISGs, such as PKR, ISG56, and IFI16, which display anti-viral functions. Conversely, high affinity IFN-α subtypes signal through ISGF3 and other factors, which activate “tunable” ISGs” such as CXCL10, IL-8, and ISG15, that induce anti-proliferative and APPLICATIONS 4 • Cellular assays (e.g. using HEK-Blue™ IFN-α/β cells) immuno-modulatory functions . IFN-α8, IFN-α10 and IFN-α14 have • ELISA. been identified as the most potent inducers of ISGs, while IFN-α1 is the weakest5. TECHNICAL SUPPORT InvivoGen USA (Toll‑Free): 888‑457‑5873 InvivoGen USA (International): +1 (858) 457‑5873 InvivoGen Europe: +33 (0) 5‑62‑71‑69‑39 www.invivogen.com InvivoGen Hong Kong: +852 3622‑3480 E‑mail: [email protected] PRODUCT DESCRIPTION Natural human IFN-α6 is induced in peripheral blood mononuclear cells upon incubation with CpG-oligonucleotides and in plasmacytoid dendritic cells upon incubation with CpG-oligonucleotides or imiquimod6. Human IFN-α6 is classified as an intermediate/strong ISG inducer5,7 with strong anti-viral activity against human metapneumovirus, mumps virus (MuV), and human immunodeficiency virus (HIV)8. Human IFN-α6 expression is IFN-α receptor-dependent as it is induced by other IFN-α subtypes upon infection with low doses of Sendai virus in vitro9. 2. Schreiber G. 2017. The molecular basis for differential type I interferon signaling. J. Biol. Chem. 292:7285-94. 3. Manry J. et al., 2011. Evolutionary genetic dissection of human interferons. J. Exp. Med. 208:2747-59. 4. Levin D. et al., 2014. Multifaceted activities of type I interferon are revealed by a receptor antagonist. Sci. Signal. 7(327). ra50. 5. Kurunganti S. et al., 2014. Production and characterization of thirteen human type-I interferon-α subtypes. Protein Expr. Purif. 103: 75-83. 6. Hillyer P. et al., 2012. Expression profiles of human interferon-alpha and interferon- lambda subtypes are ligand- and cell-dependent. Immunol. Cell. Biol. 90(8):774-83. 7. Moll H.P. et al., 2011. The differential activity of interferon-α subtypes is consistent among distinct target genes and cell types. Cytokines. 53:52-59. 8. George J. & Mattapallil J.J., 2018. Interferon-α subtypes as an adjunct therapeutic approach for human immunodeficiency virus functional cure. Front. Immunol. 9:999. 9. Zaritsky L.A. et al., 2015. Virus multiplicity of infection affects type I interferon subtype induction profiles and interferon-stimulated genes. J. Virol. 89:11534-48. PROCEDURE Preparation of stock solution (20 μg/ml): 1. Add 50 µl endotoxin-free water (provided) to 1 µg of recombinant hIFN-α. 2. Mix by pipetting. Do not vortex. 3. Use immediately or prepare aliquots of recombinant human IFN-α and store at -80 °C for 4 months. Avoid freeze-thaw cycles. 4. Further dilutions can be prepared in the appropriate aqueous buffer, such as cell culture medium containing serum. RELATED PRODUCTS Product Description Cat. Code Recombinant hIFN-α set Set of 12 hIFN-α subtypes rcyck-hifna Anti-hIFN-α-IgG Neutralizing antibody mabg-hifna-3 HEK-Blue™ IFN-α/β cells Type I IFN reporter cells hkb-ifnab LumiKine™ Xpress hIFN-α Bioluminescent ELISA kit luex-hifna TECHNICAL SUPPORT InvivoGen USA (Toll‑Free): 888‑457‑5873 InvivoGen USA (International): +1 (858) 457‑5873 InvivoGen Europe: +33 (0) 5‑62‑71‑69‑39 www.invivogen.com InvivoGen Hong Kong: +852 3622‑3480 E‑mail: [email protected].